甘油干燥保存角膜内皮细胞活性的研究

A Study on the activities of the corneal endothelial cells preserved by glycerol-desiccating

采用锥蓝 -茜素红联合染色和酶组织化学染色技术 ,对经过甘油干燥保存 1、3、6个月的猫角膜内皮细胞进行活性检测。结果显示保存不同时期的角膜内皮细胞经锥蓝 -茜素红联合染色后 ,细胞核均深度蓝染 ,蓝染率10 0 % ,细胞边界不清晰。经组织培养 7天后 ,细胞核蓝染率仍为 10 0 %。ACP、ATPase、SDH和苹果酸脱氢酶在各时期保存角膜内皮细胞中均可检测到。保存 1、3个月角膜中 ,四种酶与正常对比无明显差异 (P>0 .0 5 )。而保存6个月组 ,除 ATPase外 ,其余三种酶反应程度降低 (P<0 .0 5 )。干燥保存角膜培养 7天后 ,内皮细胞中均未再检测到酶活性 ,认为经甘油干燥保存的角膜内皮细胞不再具有活性。

To observe whether the corneal endothelial cells preserved by glycerol-desiccating have activities the cat corneal endothelial cells preserved for 1,3and 6 months were actively stained by 0 25% typan blue with1% alizarin reds respectivly At one time,four enzymes inthe corneal endothelial cells were detected by enzyme histochemical staining technology The results showed all nucleus of the endothelial cells preserved for various duration appeared heavey blue after staining by typan blue with alizarin reds The endothelial hexagonal cellular borders were not clear When cultured for 7 days,all nucleus were still heavey blue stained Under enzyme histochemical,ACP,ATPase,SDH and Malate dehydrogenase were detected in the endothelial cells preserved for various duration Compared with the fresh group,the reaction level of the cells preserved for 1 and 3 months showed no significant differences(p>0 05) While the cells preserved for 6 months presented lower level(p<0 05)except the ATPase Cultured for 7 days,there were no reaction in the all preserved cells We concluded the corneal endothelial cells preserved by glycerol-edesiccarting had no activities [WT5”HZ]